The Cell Isolation and Culture Core provides expertise and services in the design, performance and interpretation of all experiments in which cultured cells and cell lines are used. Personnel in the core have experience spanning the field of cell culture. The specific aims of the core are to 1) isolate and culture primary cells and cell lines required by each investigator for the five projects proposed in this application, 2) maintain special cell lines (e.g., stably-transfected tracheal myocytes) required for proposed investigations, 3) validate quality control for all cell cultures and cell lines used in proposed investigations, 4) maintain facilities and equipment required for these projects, and 5) develop new techniques in cell culture as required for each project, and instruct new project research personnel in proper technique. The specific services to be provided by this core are: isolation, growth and preparation of specific primary cells and cell lines, consultation on experimental design, provision and maintenance of specialized cell lines, transfection of cell lines using plasmids provided by investigators in the specific projects, and provision of data required to validate cell lines and transfected cells. The facilities and personnel involved in this core have a role in each of the scientific projects. Each project shares a need for isolation and/or culture of cells for use in experiments. There are common techniques that will have a more efficient economy of scale. In addition, common equipment needs for long-term storage of cells, and cell transfections, can be met more easily through the Core Facility. Finally, we have the necessary experience and resources available to respond and adapt to new directions of inquiry. All projects will benefit from the interaction with this Core Facility.